Genotyping and molecular investigation of plasmid-mediated carbapenem resistant clinical <i>Klebsiella pneumoniae</i> isolates in Egypt

Kholoud Baraka; Rania Abozahra; Marwa Mohammed Haggag; Sarah M Abdelhamid; Kholoud Baraka; Rania Abozahra; Marwa Mohammed Haggag; Sarah M Abdelhamid

doi:10.3934/microbiol.2023014

AIMS Microbiology

2023, Volume 9, Issue 2: 228-244. doi: 10.3934/microbiol.2023014

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Research article

Genotyping and molecular investigation of plasmid-mediated carbapenem resistant clinical Klebsiella pneumoniae isolates in Egypt

1.
Microbiology and Immunology Department, Faculty of Pharmacy, Damanhour University, El Behira, Egypt
2.
Microbiology and Immunology Department, Faculty of Pharmacy, Sinai University, Arish Campus, Sinai, Egypt

Received: 02 January 2023 Revised: 14 March 2023 Accepted: 17 March 2023 Published: 23 March 2023

Klebsiella pneumoniae is a multidrug-resistant nosocomial pathogen. Carbapenem resistance is mediated mainly by enzymes carried on transmissible plasmids causing their dissemination among other members of Enterobacteriaceae. This study aimed to molecularly detect carbapenem resistance genes in K. pneumoniae clinical isolates, genotype them using ERIC-PCR, and investigate plasmid transformation of resistant genes by using ERIC-PCR and sequencing.
Methods
Antimicrobial resistance of sixty carbapenem-resistant K. pneumoniae strains was evaluated by using the disc diffusion method. Five carbapenemases' genes were amplified by conventional PCR. Genotyping was performed using ERIC-PCR. Gene transformation was performed for the five genes to sensitive isolates. Wild and transformed isolates were genetically investigated using ERIC-PCR and sequencing.
Results
Carbapenem resistance in our isolates was associated with high resistance to all tested antibiotics. The 60 K. pneumoniae isolates were divided into 6 resistor types. The prevalence of KPC, IMP, VIM, NDM, and OXA-48 genes were 17%, 63%, 93%, 85% and 100%, respectively. Dendrogram analysis showed 57 distinct patterns, arranged in three clusters. The five genes were transformed successfully into sensitive isolates. ERIC profiles of wild and transformed isolates showed cluster A contained all the wild isolates, and cluster B contained all transformed isolates. Genetic sequences of the 5 genes reflected high genetic similarity with the GenBank reference genes before plasmid transformation; however, a distinguishable decrease of genetic similarity was observed after transformation.
Conclusion
Plasmid-mediated carbapenem resistance in K. pneumoniae and its dissemination among different strains is a real threat to public health.
- Klebsiella,
- Egypt,
- carbapenem,
- genotyping,
- plasmid
Citation: Kholoud Baraka, Rania Abozahra, Marwa Mohammed Haggag, Sarah M Abdelhamid. Genotyping and molecular investigation of plasmid-mediated carbapenem resistant clinical Klebsiella pneumoniae isolates in Egypt[J]. AIMS Microbiology, 2023, 9(2): 228-244. doi: 10.3934/microbiol.2023014

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Abstract

Klebsiella pneumoniae is a multidrug-resistant nosocomial pathogen. Carbapenem resistance is mediated mainly by enzymes carried on transmissible plasmids causing their dissemination among other members of Enterobacteriaceae. This study aimed to molecularly detect carbapenem resistance genes in K. pneumoniae clinical isolates, genotype them using ERIC-PCR, and investigate plasmid transformation of resistant genes by using ERIC-PCR and sequencing.

Methods

Antimicrobial resistance of sixty carbapenem-resistant K. pneumoniae strains was evaluated by using the disc diffusion method. Five carbapenemases' genes were amplified by conventional PCR. Genotyping was performed using ERIC-PCR. Gene transformation was performed for the five genes to sensitive isolates. Wild and transformed isolates were genetically investigated using ERIC-PCR and sequencing.

Results

Carbapenem resistance in our isolates was associated with high resistance to all tested antibiotics. The 60 K. pneumoniae isolates were divided into 6 resistor types. The prevalence of KPC, IMP, VIM, NDM, and OXA-48 genes were 17%, 63%, 93%, 85% and 100%, respectively. Dendrogram analysis showed 57 distinct patterns, arranged in three clusters. The five genes were transformed successfully into sensitive isolates. ERIC profiles of wild and transformed isolates showed cluster A contained all the wild isolates, and cluster B contained all transformed isolates. Genetic sequences of the 5 genes reflected high genetic similarity with the GenBank reference genes before plasmid transformation; however, a distinguishable decrease of genetic similarity was observed after transformation.

Conclusion

Plasmid-mediated carbapenem resistance in K. pneumoniae and its dissemination among different strains is a real threat to public health.

Conflict of interest

The authors declare no conflict of interest.

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